The HPV transcriptome in HPV16 positive cell lines

Abstract

Background Infections with high-risk human papillomaviruses (HPV), mainly HPV type 16, can cause malignant transformation of the human cervical epithelium and cervical cancer (CxCa). Very little is known about the quantitative expression of the various HPV16 transcripts in frequently used cervical cancer cell lines. Methods We have quantitatively analysed the viral transcriptome in the HPV16-transformed cell lines SiHa, CaSki, MRI-H196, MRI-H186, HPK-IA and C3. We used a nucleic acid sequence-based amplification (NASBA)–Luminex hybridisation assay quantifying spliced and unspliced HPV16 transcripts. Results The cell lines differed in their qualitative and quantitative expression of viral transcripts depending on the physical HPV genome status. In SiHa, we found no transcripts containing the splice acceptor at nucleotide 3358 or downstream sequences. In CaSki cells virtually all viral transcripts were detected but with a reduced quantity of late transcripts. We further found that the tumorigenic phenotype of late passage HPK-IA cells may not be mediated through changes in HPV expression. In MRI-H186, HPK-IA and C3 cells very high levels of full-length early transcripts ending at the early polyadenylation signal were found. MRI-H196 expressed L1 full-length (fl) but no E5 fl RNA suggesting the presence of integrated HPV16 genomes with a disrupted E2 region and rearranged L1 DNA sequence. Conclusion Quantitative expression changes of HPV16 transcript markers correlate with the physical state of the HPV genome in HPV16 positive cervical cancer cell lines.